Species-specific method for identifying infecticity of eimeria species

摘要

Unique methods are disclosed to determine the relative infectivity of each of multiple species of Eimeria in a multivalent live vaccine. Genomic deoxyribonucleic acid (DNA) is extracted from intestinal epithelia and mucosa of vaccinated birds and amplified using the polymerase chain reaction (PCR). The reaction allows efficient amplification of fragments from every small subunit ribosomal RNA (ssrRNA) gene in the target pool. Unique species-specific hybridization probes complimentary to sequences within the heterogeneous pool of PCR products are used to quantitate the relative levels of each of the Eimeria species in the infected hosts. The same unique probes are used to identify and quantitate species specific ssrRNA sequences in total RNA extracted from intestinal epithelia and mucosa of vaccinated chickens. The same unique probes are also used to identify and quantitate species specific ssrRNA sequences in genomic DNA isolated from mixed oocyst population purified from fecal material obtained from vaccinated chickens.